Metabolism of exogenous arachidonic acid by murine macrophage-like tumor cell lines.
Murine macrophage-like cell lines, J774.2, P388D1, RAW264.7 and PU-5-1R, were incubated with exogenous arachidonic acid (AA). The major metabolites were identified by comigration with known standards in TLC and HPLC and by characteristic behavior following reduction. During a 30 min incubation J774.2 cells metabolized exogenous 14C-AA (10 microM) to PGE2 (14.8%), 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT) (13.0%), thromboxane B2 (TXB2) (7.4%), PGD2 (4.4%) and PGF2 alpha (3.0%). The remainder was incorporated into phospholipids (39.0%), triglycerides (6.1%), and as yet unidentified metabolites (8.2%). No PGF1 alpha was found. Metabolism of exogenous AA was rapid, being less than 90% completed at 3.5 min. Metabolism of exogenous AA is not increased by the simultaneous addition of macrophage stimuli including the cation ionophore A-23187, particulate phagocytic stimuli and endotoxin. The synthesis of cyclooxygenase products was inhibited by low doses of indomethacin (ID50=0.6 microM) while the synthesis of TXB2 and HHT was selectively inhibited by benzylimidazole (ID50=9.5 microM). Identification of a probable lipoxygenase product is being pursued. The synthesis of this product is not inhibited by indomethacin and migrates with an Rf value close to 5,12-diHETE in TLC. P388D1 and RAW264.7 cells metabolize exogenous AA to the same products as J774.2, but in different proportions, while PU-5-1R does not produce cyclooxygenase metabolites to any appreciable extent.
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D002460:Cell Line; D016861:Cyclooxygenase Inhibitors; D015237:Dinoprost; D015232:Dinoprostone; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006880:Hydroxy Acids / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D007213:Indomethacin / Q000494:pharmacology; D007700:Kinetics; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D009374:Neoplasms, Experimental / Q000378:metabolism*; D015230:Prostaglandin D2; D011457:Prostaglandins D / Q000378:metabolism; D011458:Prostaglandins E / Q000378:metabolism; D011460:Prostaglandins F / Q000378:metabolism; D013929:Thromboxane B2 / Q000378:metabolism; D013930:Thromboxane-A Synthase / Q000037:antagonists & inhibitors
D001095:Arachidonic Acids; D016861:Cyclooxygenase Inhibitors; D005231:Fatty Acids, Unsaturated; D006880:Hydroxy Acids; D007093:Imidazoles; D011457:Prostaglandins D; D011458:Prostaglandins E; D011460:Prostaglandins F; D016718:Arachidonic Acid; C017062:1-benzylimidazole; C008033:12-hydroxy-5,8,10-heptadecatrienoic acid; D013929:Thromboxane B2; D015237:Dinoprost; D013930:Thromboxane-A Synthase; D015232:Dinoprostone; D015230:Prostaglandin D2; D007213:Indomethacin